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Mohit Parekh, Gianni Salvalaio, Stefano Ferrari, Alessandro Ruzza, Marie-Claude Amoureux, Denis Fortier, Diego Ponzin; Synthetic media for replacement of serum based conventional organ culture corneal preservation system. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):1114.
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© ARVO (1962-2015); The Authors (2016-present)
To evaluate the efficacy of a new synthetic medium (Cornea Syn, Eurobio, France) and compare it with conventional organ culture medium (Cornea Max, Eurobio, France).
Cornea Syn is Iscove based, serum free and completed with recombinant factors needed for maintaining cornea healthy in organ culture. Seven pairs of human donor corneas were evaluated using Cornea Syn against the traditional serum based Cornea Max. Each cornea from the same donor was preserved in Cornea Cold or Cornea Prep II for 48 hours at RT [phase I], transferred and preserved in Cornea Syn or Cornea Max for 28 days at 31oC [phase II] followed by preservation in Cornea Trans or Cornea Jet for 4 days at RT [phase III]. Parameters such as thickness, transparency, viable endothelial cell density (VECD), morphology and overall quality were used to determine the quality of the cornea. Trypan blue for cell mortality; Lactic acid production in the media; Alizarin red for hexagonality; ZO-1, p63 and alpha SMA immunostaining for tight junctions, limbal and smooth muscle actin in the stroma respectively on histological sections; Hematoxylin Eosin staining for corneal integrity and TdT dNTP kit for cell apoptosis, were used for analytical study.
Thickness, transparency and overall quality showed statistical significance (p<0.05) showing better results with Cornea Syn at phase I however, none of the other parameters showed any significance at any stage. Cornea Syn showed statistically significantly lower (p<0.05) production of lactic acid as compared to Cornea Max however, no statistical significance was observed in other phases. Alizarin red showed partial preservation of hexagonality as the morphology deteriorated to some extent in both series. Immunostaining showed expression of tight junctions of the endothelium, preservation of the limbal region of the epithelium and muscle fibres in the stroma. Hematoxylin Eosin staining showed presence of all the corneal layers. No apoptosis was observed in any preserved corneas.
Cornea Synthetic series is comparable to the conventional serum based media which helps to preserve the corneal integrity and metabolism active. Cornea Syn is a safe and reliable media for preservation of human donor corneal tissues at 31oC and has an advantage of having no animal or animal derived components.
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